Digenome-seq

Cas9-Digested Whole-Genome Sequencing

Digenome-seq was designed to profile genome-wide Cas9 off-target effects (Kim et al., 2015). A multiplexed version has also been published (Kim et al., 2016). It belongs to a family of methods, including HTGTS (Chiarle et al., 2011), LAM-HTGTS (Hu et al., 2016), and Guide-seq (Kim et al., 2015), which are aimed at detecting off-target effects of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 and other RNA-guided engineered nucleases (RGENs).

This method detects off-target mutations, induced by RGENs in a bulk population of cells, by sequencing in vitro nuclease-digested genomes (digenomes). These digests should produce many DNA fragments with identical 5_ ends, which are vertically aligned at cleavage sites.

Advantages:

Relies on DNA cleavage rather than binding

Performed in a genomic context and captures sites with a DNA/RNA bulge

Related

Edit: Digenome-seq

History: Digenome-seq

Digenome-seq was designed to profile genome-wide Cas9 off-target effects (Kim et al., 2015). A multiplexed version has also been published (Kim et al., 2016). It belongs to a family of methods, including HTGTS (Chiarle et al., 2011), LAM-HTGTS (Hu et al., 2016), and Guide-seq (Kim et al., 2015), which are aimed at detecting off-target effects of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 and other RNA-guided engineered nucleases (RGENs).

This method detects off-target mutations, induced by RGENs in a bulk population of cells, by sequencing in vitro nuclease-digested genomes (digenomes). These digests should produce many DNA fragments with identical 5_ ends, which are vertically aligned at cleavage sites.

Advantages:

Relies on DNA cleavage rather than binding

Performed in a genomic context and captures sites with a DNA/RNA bulge

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